Generally Recognized As Safe Research Articles

Montmorillonite nanosheets as an adjuvant eliciting potent cellular immunity

The experience of fighting COVID-19 shows the growing importance of vaccine-induced cellular immunity, whereas aluminum adjuvants fail to meet this medical requirement. Herein, with very simple processing, we prepared a narrow lateral size of monolayer clay nanosheets from montmorillonite, a FDA-approved generally recognized as safe (GARS) material. The nanosheets adsorbed antigen ovalbumin (OVA) through hydrophilic and electrostatic interactions, while utilizing nanosheets to expose more octahedral Al-OH and tetrahedral Si-OH residues to complete the adsorption of mannose (Man) targeting antigen presentation cells, leading to the production of nanosheet vaccines. In additional, the lyophilization process did not significantly affect the loading of OVA and Man on the nanosheets. The obtained nanosheet vaccines induced a strong cellular immune response in addition to a potent humoral immune response. Inspired by the results, the prophylactic nanosheet adjuvant vaccine against Epstein-Barr virus (EBV) was developed, which induced a 13-fold increase in EBV-neutralizing antibody titers compared to aluminum adjuvant vaccine, and efficiently blocked EBV infection of target cells. Based on the simple formulation process, remarkable immune response effect and excellent safety profile, montmorillonite nanosheets have the potential to replace aluminum adjuvant as a vaccine platform for inducing effective immune responses.

Effectiveness of GRAS substances for managing leaf diseases in brinjal (<em>Solanum melongena</em> L)

Leaf diseases in brinjal (Solanum melongena L.) are common and drastically lower its yield. As synthetic fungicides are hazardous to the environment and human health, non-fungicidal approaches to foliar disease control are being adopted progressively. The current investigation assessed the effectiveness of Generally Recognized as Safe (GRAS) substances in the management of leaf diseases in brinjal. The efficacy of the selected GRAS substances, including 400 μM of salicylic acid, 0.1% of sodium bicarbonate, and botanical extracts such as 200 ppm of garlic clove extract, 200 ppm of neem extract and 200 ppm of eucalyptus extract, were assessed. Six treatments and four replicates were employed in a completely randomized design for this present study. The natural occurrence of leaf diseases was taken into account in the experiment. Disease incidence (%), disease severity (%), number of leaves per plant, plant height (cm) and number of branches per plant were recorded seven weeks after transplanting. Analysis of variance was used to assess the significance of treatment effects. The findings confirmed that the exogenous spraying of salicylic acid and garlic clove extract significantly (P<0.05) decreased the disease severity and incidence, with statistically equivalent efficiency for both substances. Moreover, number of branches per brinjal plant (3.3), plant height (51.4 cm) and number of leaves per brinjal plant (31.5) were significantly (P<0.05) enhanced by salicylic acid compared to control. Hence, spraying garlic clove extract and salicylic acid may therefore be an environmentally potential means of managing brinjal leaf diseases during the vegetative growth stage.

Pain and Inflammation Management: Part-II Clinical Investigation of a Topical Ayurvedic Cream called HerboCare or HerboJointTM

Background: Joint pain and chronic inflammation pose significant health challenges, particularly among the elderly population. Current treatments often offer only temporary relief and are associated with potential side effects. Ayurvedic botanical herbs and oils, known for their phytochemical-rich, anti-inflammatory properties, hold promise in addressing these issues.Objective: This study aimed to investigate the clinical efficacy of HerboCare orHerboJoint™, a topical Ayurvedic cream, in alleviating muscle discomfort, inflammation, and joint pain, along with its impact on inflammatory cytokines, particularly TNF-α on human subjects. While HerboCare or HerboJoint had a very low content of Menthol, HerboJoint-Plus (not reported in this study) was formulated with a much higher content of USP grade Menthol.Methods: HerboCareor HerboJoint™ cream, formulated with essential oils from Cymbopogon citratus, Hedychium spicatum, Zanthoxylum alatum, and Menthol (Mentha arvensis), was evaluated in two sets of clinical trials over a 90-day period. Patients with joint pain were divided into different treatment groups, and their pain, swelling, stiffness, and inflammatory marker TNF-α were assessed.Results: In the first set of trials, HerboCare orHerboJoint™ significantly reduced joint pain (29%), joint swelling (31%), and joint stiffness (60%) in patients. In the second set, TNF-α levels were reduced by 38% in the HerboCare orHerboJoint™-only group, 63% in the HerboCare orHerboJoint™ and prescribed formulation group, and 71% in the group receiving only the prescribed formulation. These results underscored the cream's efficacy in mitigating inflammation.Conclusion: HerboCare orHerboJoint™, formulated with Generally Recognized as Safe (GRAS)-affirmed botanical ingredients, demonstrates clinical effectiveness in relieving joint pain and inflammation. This Ayurvedic approach offers a safe and cost-effective alternative for managing these debilitating conditions.Keywords: Muscular discomfort,Joint Pain, Essential Oils; Menthol; Inflammatory Cytokines; Clinical Study

Enhancement of dieckol extraction yield from Ecklonia cava through optimization of major variables in generally recognized as safe solvent-based process

Ecklonia cava (EC), an edible brown macroalga abundant in intertidal areas of East Asia (Korea, Japan, and China), contains high-value bioactive compounds such as dieckol, which has antifungal, anti-inflammatory, antitumor, and antihyperlipidemic activities. However, no studies have been reported on the utilization of EC as a biorefinery feedstock, and the design of a more economical and high-yield process is required for the utilization of dieckol for the human healthcare industry. In this study, we designed a bioprocess for the high-yield recovery of dieckol from EC in a generally recognized as safe (GRAS) solvent to facilitate its application in the food and healthcare industries. Preliminary studies identified ethanol as an efficient solvent with the highest dieckol extraction yield (2.9 mg/g biomass). In order to maximize the recovery of dieckol from EC, the major extraction variables (solvent concentration, reaction temperature, and reaction time) were optimized based on statistical methods. Based on the predictive model, the numerical optimization determined that the solution with the highest dieckol content per weight of extract (62.6 vol% ethanol concentration, 54.2°C temperature, 13.2 min) was the optimal extraction condition. Under the determined conditions, the dieckol yield from EC achieved 6.4 mg dieckol/g EC (95.5% agreement with the predicted value). The designed process offers several advantages, including improving the utilization feasibility of EC, utilizing GRAS solvents with potential human applications, short extraction time (13.2 min), maximized process yield, and the highest dieckol recovery compared to previous reports.

Antioxidant properties of wheat bran FOs prepared by Bacillus amyloliquefaciensIT‐45 fermentation

SummaryAs functional oligosaccharides, feruloyl oligosaccharides (FOs) exhibit the physiological activities of both ferulic acid and oligosaccharides. In this study, FOs were prepared by fermenting wheat bran with the generally recognised as safe (GRAS) bacterium Bacillus amyloliquefaciens IT‐45. On the fourth day of fermentation, the activities of xylanase and cellulase reached 447.67 and 33.41 U/mL, respectively. UV, FTIR, and HPLC were used to characterise the structure properties of wheat bran FOs from B. amyloliquefaciens IT‐45 fermentation. The monosaccharide compositions of FOs were mainly composed of xylose and arabinose. Under the optimal conditions of fermentation time of 4 days, fermentation temperature of 25 °C, initial pH of 6.5 and inoculation amount of 11% (v/v), the maximum yield of FOs in the broth was 0.6397 mM. In vitro antioxidant activity analysis showed that FOs prepared by B. amyloliquefaciens fermentation had good scavenging ability against ABTS, DPPH and hydroxyl radicals, demonstrating the application potential. The results provided a relatively environmentally friendly and food‐safety way to prepare feruloyl oligosaccharides.

Increasing shelf life of millets by developing bio-nanocomposite films (bioplastic) as an alternative to conventional plastic

Millets are considered as most nutritional grain among all. Hence it becomes immense important to prevent and protect such grains from being deteriorated. Spoilage is mostly observed due to bacterial, fungal, or due to some climatic changes. Such spoilage cannot be stopped completely but can be minimized by increasing its shelf life. Development of metal-oxide based bio-nanocomposite film has revolutionized various sectors from food packaging to agriculture. Zinc oxide nanoparticles is considered as GRAS (Generally Recognized As Safe) among all metal-oxides, and hence introduced in various applications. These ZnO NPs can be combined with natural polymer forming bio-nanocomposite films (bioplastic) and therefore used in mulching and food packaging industry. Mulching improves crop yield, requires less water, makes less usage of pesticide, and promote proper growth of plants. Till date, conventional plastic made of polyethylene units are used which causes hazardous effect on environment, hence it become immense important to replace it with a biodegradable and affordable bioplastic. This study involves biosynthesis of ZnO NPs using plant (Cassia fistula) extract and combining it with the natural biopolymer, giving rise to bio-nanocomposite film. These films would show antibacterial activity against pathogen causing spoilage in millets and thus increasing their shelf-life extension. Also, the minimum amount of ZnO NPs required to show antibacterial activity would be determined.

Improved production of lactiplantibacillus plantarum RO30 exopolysaccharide (REPS) by optimization of process parameters through statistical experimental designs

BackgroundIn investigating of (exopolysaccharide) EPS from unconventional sources, lactic acid bacteria have a vital role due to their generally recognized as safe (GRAS) status. EPSs have diverse applications such as drug delivery, antimicrobial activity, surgical implants, and many more in many sectors. Despite being important, the main hindrance to the commercial application of these significant biopolymers is low productivity. Therefore, this study primarily focuses on optimizing physio-chemical conditions to maximize the previously produced EPS from probiotic Lactiplantibacillus plantarum RO30 (L. plantarum RO30) using one factor at a time (OFAT) and method Response Surface Methodology (RSM).ResultsThe EPS obtained from L. plantarum RO30 named REPS. The medium formulation for REPS production using the OFAT method revealed that sucrose (20 g/L, beef extract (25 g/L), and ammonium sulfate at 4 g/L concentration were the optimum carbon, organic and inorganic nitrogen sources, and REPS yield was increased up to 9.11 ± 0.51 g/L. RSM experiments revealed that, a greatly significant quadratic polynomial attained from the Central Composite Design (CCD) model was fruitful for specifying the most favorable cultural conditions that have significant consequences on REPS yield. The maximal amount of REPS (10.32 g/L) was formed by: sucrose (40 g/L), beef extract (25 g/L), pH (5.5), incubation temperature (30 °C), and incubation period (72 h). A high closeness was obtained between the predicted and experimental values and it displayed the efficiency of the RSM.ConclusionThis study was conducted to reinforce REPS production in the probiotic LAB L. plantarum RO30 by utilizing various experimental parameters. The maximum REPS yield of 10.32 g/L was attained under the circumstances optimized in the study.

Biotransformation of Sclareol by a Fungal Endophyte of Salvia sclarea.

Microbial endophytes are known as versatile producers of useful metabolites, which have extensive applications in pharmacy, fragrance, agriculture and food. This study aims to screen sclareol-biotransforming microorganisms from Salvia sclarea, an untapped source of diverse endophytes. In this study, 50 culturable endophytes were isolated from S. sclarea grown in Xinjiang using sclareol as the sole carbon source and screened for their potential to transform sclareol into analogues. A fungal endophyte, identified as the generally recognized as safe (GRAS) strain Aspergillus tubingensis, can produce labd-14-ene-3β,8α,13β-triol and 8α,13β-dihydroxylabd-14-en-3-one from sclareol, involving hydroxylation and carbonylation at the C3 site. Structures of the two metabolites were elucidated by HR-ESI-MS and NMR analysis. S. sclarea was proven to be a good source of endophytes that are prospective producers of secondary metabolites with valuable chemical and biological properties. This study is the first report regarding the isolation of endophytes from S. sclarea.

Enzymatic and antibacterial activity of the recombinant endolysin PVP-SE1gp146 expressed in Hansenula polymorpha

Antibiotic resistance, a major global concern, highlights the need for discovering alternative therapies. Recently, endolysins have garnered attention as antibacterial tools with a lower resistance development rate compared to conventional antibiotics, and their production in various expression hosts holds significance. Given its generally recognized as safe (GRAS) status and other advantages, Hansenula polymorpha offers a promising host for endolysin production. PVP-SE1gp146 originates from the Salmonella Enteritidis-specific phage PVP-SE1, which has been previously characterized. We inserted the PVP-SE1gp146 coding gene into the H. polymorpha expression vector pHIPX4. The resulting recombinant, pHIPX4-PVP-SE1gp146, was then introduced into H. polymorpha NCYC495 to facilitate the production of the endolysin PVP-SE1gp146. The expression level of the PVP-SE1gp146 protein was assessed, and it was determined to be approximately 43 mg/l of yeast culture medium. The enzymatic (muralytic) activity of this endolysin was also evaluated, corresponding to the version produced by the E. coli Bl21 strain. The endolysin exhibited admissible antibacterial activity against several gram-negative species, including P. aeruginosa, E. coli, and A. baumannii, while showing an almost negligible impact on K. pneumoniae. Endolysin production within GRAS-approved hosts holds potential for combating antibiotic-resistant bacteria. Challenges involve optimizing concentrations, targeting gram-negative species and improving attachment to bacterial cell walls. Addressing these issues requires dedicated research in endolysin engineering and a comprehensive evaluation of their production in diverse expression hosts.

60 Aafco Approval and Regulatory Process for Novel Ingredients

Abstract In this presentation, we will explore different pathways in the regulatory approval process. There are three routes that individuals or companies can take to gain approval for an ingredient in a target species. The most common and rigorous of the three is the AAFCO Ingredient Definition process, which takes at least 18 months from start to finish. This process involves a six-month FDA review period of safety data generated in a target species, gathered after feeding a range of inclusions of the target ingredient. Additionally, manufacturing and analytical data are required for this process. After FDA review, there is a minimum of one year for review by the AAFCO ingredient definition committee and general membership. The second regulatory route we will discuss is Generally Recognized as Safe (GRAS). This route can be shorter but will require similar safety data and an assessment by an expert panel is recommended. FDA review of the GRAS panel findings is necessary for listing in the AAFCO original publication as GRAS. Finally, we will discuss the GRAS self-determined pathway. This does not require FDA review but entails state-by-state registration of products. This pathway will still require safety, analytical and efficacy data to support the product registration with each state.

A sweeter future: Using protein language models for exploring sweeter brazzein homologs.

With growing concerns over the health impact of sugar, brazzein offers a viable alternative due to its sweetness, thermostability, and low risk profile. Here, we demonstrated the ability of protein language models to design new brazzein homologs with improved thermostability and potentially higher sweetness, resulting in new diverse optimized amino acid sequences that improve structural and functional features beyond what conventional methods could achieve. This innovative approach resulted in the identification of unexpected mutations, thereby generating new possibilities for protein engineering. To facilitate the characterization of the brazzein mutants, a simplified procedure was developed for expressing and analyzing related proteins. This process involved an efficient purification method using Lactococcus lactis (L. lactis), a generally recognized as safe (GRAS) bacterium, as well as taste receptor assays to evaluate sweetness. The study successfully demonstrated the potential of computational design in producing a more heat-resistant and potentially more palatable brazzein variant, V23.

In Vitro Evaluation of the Adsorption Efficacy of Biochar Materials on Aflatoxin B1, Ochratoxin A, and Zearalenone

Mycotoxin sequestration materials are important tools to reduce mycotoxin illness and enable proper handling of mycotoxin-contaminated commodities. Three food-grade bentonite clays and four generally recognized as safe (GRAS) charcoal/biochar carbon materials that are marketed as feed additives and supplements were evaluated for their ability to sequester the mycotoxins aflatoxin B1, ochratoxin A, and zearalenone. The surface area of the clays varied between 32.1 to 51.4 mg2/g, and the surface area of the carbon-based materials varied from 1.7 to 1735 mg2/g. In vitro, gastric fluid studies indicated that certain pine biochar and activated coconut charcoal could sequester high amounts (85+%) of the mycotoxins at 1 ppm levels or below. However, some biochar materials with lower surface area properties lacked binding capacity. The coconut shell charcoal and pine biochar utilize agricultural waste products in a manner that significantly reduces carbon emissions and provides valuable materials to minimize exposure to toxins found in food and feed.

Fabric phase sorptive extraction as a sustainable sample preparation procedure to determine synthetic musks in water

A rapid sample preparation fabric phase sorptive extraction (FPSE) methodology followed by gas chromatography coupled to tandem mass spectrometry is proposed for the first time to determine 12 synthetic musks including nitro-, polycyclic and macrocyclic musk in environmental water and wastewater. An asymmetric screening design of experiments was used to evaluate the influence of nine parameters affecting FPSE in only 16 experiments. The factors included the sol–gel sorbent coating, stirring mode (both extraction and desorption), extraction and desorption time, salting-out effect, sample volume, type of solvent, and desorption solvent volume. The selected conditions imply the use of the sol–gel PDMS sorbent coating, 15 min extraction by ultrasound energy employing 10 mL of sample and 0.5 mL of ethanol as desorption solvent during 5 min. Accuracy and precision were assessed in different real water samples. Matrix effects were evaluated performing recovery studies in several aqueous matrices: river, sea, spring, laundry washing place, and wastewater at different concentration levels (0.1, 2, 10 µg L−1) demonstrating method accuracy (results ranged between 82 and 110%) and good precision (relative standard deviation, RSD < 12% in all cases). Limits of detection (LODs) were below 8 ng L−1 for the analyzed synthetic musks. As extraction was quantitative (exhaustive extraction), calibration was carried out using solvent standards without the need of repeating the extraction step, which is required in most microextraction methods, improving sample throughput, and reducing costs. Finally, the analysis of real contaminated samples revealed the presence of 11 out of the 12 target synthetic musks at concentrations up to 30 µg L−1, three of them banned in cosmetic regulation (musk tibetene, ambrette, and moskene). The developed FPSE based methodology uses low volume (0.5 mL) of a Generally Recognized as Safe (GRAS) solvent (ethanol), as well as minimum energy and time consumption, demonstrating to be a sustainable alternative to detect these microcontaminants in environmental waters and wastewaters.

Antibacterial Activity of Zinc Oxide Nanoparticles Loaded with Essential Oils.

One major problem with the overuse of antibiotics is that the microorganisms acquire resistance; thus the dose must be increased unsustainably. To overcome this problem, researchers from around the world are actively investigating new types of antimicrobials. Zinc oxide (ZnO) nanoparticles (NPs) have been proven to exhibit strong antimicrobial effects; moreover, the Food and Drugs Administration (FDA) considers ZnO as GRAS (generally recognized as safe). Many essential oils have antimicrobial activity and their components do not generate resistance over time. One of the drawbacks is the high volatility of some components, which diminishes the antimicrobial action as they are eliminated. The combination of ZnO NPs and essential oils can synergistically produce a stronger antimicrobial effect, and some of the volatile compounds can be retained on the nanoparticles' surface, ensuring a better-lasting antimicrobial effect. The samples were characterized with X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), ultraviolet-visible spectroscopy (UV-Vis), and thermal analysis (TG-DSC) coupled with analysis of evolved gases using FTIR. The ZnO NPs, with a size of ~35 nm, exhibited a loading between 1.44% and 15.62%-the lower values were specific for limonene-containing oils (e.g., orange, grapefruit, bergamot, or limette), while high values were obtained from cinnamon, minzol, thyme, citronella, and lavender oils-highlighting differences among non-polar terpenes and alcohol or aldehyde derivatives. The antibacterial assay indicated the existence of a synergic action among components and a high dependency on the percentage of loaded oil. Loaded nanoparticles offer immense potential for the development of materials with specific applications, such as wound dressings or food packaging. These nanoparticles can be utilized in scenarios where burst delivery is desired or when prolonged antibacterial activity is sought.

Therapeutic Effects of Geranium Oil in MPTP-Induced Parkinsonian Mouse Model.

Parkinson's disease (PD) is an incurable neurodegenerative disease characterized by motor and non-motor disabilities resulting from neuronal cell death in the substantia nigra and striatum. Microglial activation and oxidative stress are two of the primary mechanisms driving that neuronal death. Here, we evaluated the effects of geranium oil on 1-methyl-4-phenyl-1,2,3,6-tetra-hydropyridine (MPTP) mouse model for PD, on microglial activation, and oxidative stress. We demonstrate that oral treatment with geranium oil improved motor performance in this model. The therapeutic effects of geranium oil were observed as a significant increase in rotarod latency and distance among the mice treated with geranium oil, as compared to vehicle-treated MPTP mice. Geranium oil also prevented dopaminergic neuron death in the substantia nigra of the treated mice. These therapeutic effects can be partially attributed to the antioxidant and anti-inflammatory properties of geranium oil, which were observed as attenuated accumulation of reactive oxygen species and inhibition of the secretion of proinflammatory cytokines from geranium oil-treated activated microglial cells. A repeated-dose oral toxicity study showed that geranium oil is not toxic to mice. In light of that finding and since geranium oil is defined by the FDA as generally recognized as safe (GRAS), we do not foresee any toxicity problems in the future and suggest that geranium oil may be a safe and effective oral treatment for PD. Since the MPTP model is only one of the preclinical models for PD, further studies are needed to confirm that geranium oil can be used to prevent or treat PD.

REGULATION OF FUNCTIONAL FOODS IN UKRAINE AND THE WORLD. PROSPECTS FOR THE USE OF POSTBIOTICS AS FUNCTIONAL INGREDIENTS

Functional food allows to individualize the characteristics and necessities of each person. Over the past few decades, the demand for products that have a positive impact on human health has exploded worldwide. The market for functional foods in developed countries is estimated at USD 300 billion, which makes this sector attractive for investment. Accordingly, the production of functional foods and ingredients needs to be regulated at the state level. There is no harmonization in the definitions of functional foods and their classification at both the global and state levels. Despite the fact that the concept and terminology of functional food and the means of achieving it is well-established, the term “functional food” is often absent in the legislative acts of countries that actively produce functional foods, and the term “health-related food” is used instead. As a rule, health-related foods are subject to quality and safety requirements that are identical to those of conventional food products. Exceptions are regulations that establish rules for labeling FFPs and statements regarding their physiological effects or the content of a particular ingredient that may affect physiological aspects. In Europe and the USA, much attention is paid to the safety of “health-related food” and the novel food products. European Food Safety Authority and the US Food and Drug Administration require sponsors to submit information on the composition of multiple batches of a product to support safety evaluations for novel foods and Generally Recognized as Safe (GRAS) ingredients. In Ukraine, the legal status of the terms “functional food product” and dietary supplement was regulated by Law of Ukraine 771 “On Basic Principles and Requirements for Food Safety and Quality”, but in the version of this law dated 16.01.2020, this term was excluded, the term “dietary supplement” was edited and the term “novel food product or ingredient” was included, which is related to the process of harmonization of Ukrainian and European legislation in the food sector. In 2020, the Ministry of Health of Ukraine approved Order No. 1145 “On Approval of the Requirements for Nutrition Claims and Health Claims for Food Products”, which regulates the following concepts: claims and conditions for their use; health claims allowed to be used in food labeling and advertising, except for claims about reducing the risk of diseases and claims related to the development and health of children; claims about reducing the risk of diseases and claims related to the development and health of children. Unfortunately, Ukrainian regulations do not contain claims and conditions for the use of claims about pro- and prebiotics that have GRAS status worldwide. Particular attention should be paid to the possibility of using postbiotics as functional food ingredients with immunological activity.

Anti-biofilm activity of essential oils in fruit and vegetable: A systematic review

Biofilm formation is one of the most serious threats in the food industry, mostly due to its contribution to the survival of spoiling and pathogenic microorganisms. Essential oils (EOs) are classified as secondary metabolites of plants which are generally recognized as safe (GRAS) and are typically mixtures of numerous components, and the monoterpenes constitute about 90% of their composition. The worldwide increase in foodborne outbreaks associated with fresh produce made us conduct a systematic review on in vitro application of EOs to remove biofilms in fruit and vegetables. Databases such as Scopus, PubMed and Web of Science were probed for relevant studies. The search criteria was restricted to published papers in English language up to August 2022. A total of 299 studies were extracted and after removing irrelevant papers, 17 papers were included in this review. Results indicated the effectiveness of EOs and their exposure time with fresh products on the breakdown of biofilms. Among the findings, only a part of a study indicated no effect of EOs (Citrus sinensis, Citrus aurantifolia, Coffea arabica and Vitex agnus-castus) against Erwinia amylovora biofilms in pear. Moreover, EOs were shown to act as pesticides and were reported to be effective against biofilm formation at pre-harvest stages. Most of the studied EOs showed strong effect on reducing bacterial burden without affecting the sensory score of fruits and vegetables; however, it is suggested that more attention should be paid to sensory evaluation of treated samples in future studies. These data underline the need for conducting more research on novel essential oils (or combinations) and characterization of them as natural antibiofilm agent.

Antifungal and Antiaflatoxigenic Activities of Massoia Essential Oil and C10 Massoia Lactone against Aflatoxin-Producing Aspergillus flavus.

Fungal infection and mycotoxin contamination are major hazards to the safe storage and distribution of foods and feeds consumed by humans and livestock. This study investigated the antifungal and antiaflatoxigenic activities of massoia essential oil (MEO) and its major constituent, C10 massoia lactone (C10), against aflatoxin B (AFB)-producing Aspergillus flavus ATCC 22546. Their antifungal activities were evaluated using a disc diffusion assay, agar dilution method, and a mycelial growth inhibition assay with the AFB analysis using liquid chromatography triple quadrupole mass spectrometry. MEO and C10 exhibited similar antifungal and antiaflatoxigenic activities against A. flavus. C10 was a primary constituent in MEO and represented up to 45.1% of total peak areas analyzed by gas chromatography-mass spectrometry, indicating that C10 is a major compound contributing to the antifungal and antiaflatoxigenic activities of MEO. Interestingly, these two materials increased AFB production in A. flavus by upregulating the expression of most genes related to AFB biosynthesis by 3- to 60-fold. Overall, MEO and C10 could be suitable candidates as natural preservatives to control fungal infection and mycotoxin contamination in foods and feeds as Generally Recognized As Safe (GRAS) in the Flavor and Extract Manufacturers Association of the United States (FEMA), and MEO is a more suitable substance than C10 because of its wider range of uses and higher allowed concentration than C10.

Out of balance: conflicts of interest persist in food chemicals determined to be generally recognized as safe

Manufacturers of chemicals added to food are responsible for determining that the use of their products is safe. There are two major legal definitions of chemicals in food: (1) food additives which includes ingredients and chemicals indirectly entering food from packaging and processing equipment, and (2) generally recognized as safe (GRAS) substances mostly used as ingredients. The law requires food additives to undergo approval by the U.S. Food and Drug Administration (FDA) before they are sold, but it GRAS substances are exempted from pre-market approval. In 1997, FDA created a voluntary program for manufacturers to submit their chemical’s safety determination in the form of a GRAS notice to the agency. Manufacturers make GRAS determinations regardless of whether they voluntarily submit a notice to FDA for review. They rely on their own employees, the employee of a hired consulting firm or a panel of experts, known as GRAS panel, to review the safety information. Because this process determines whether a chemical is safe for use in food, conflicts of interest and biases need to be avoided or minimized to credibly ensure food is safe. Recently, FDA has published guidance for industry on best practices to convene GRAS panels to manage conflicts of interest and reduce biases that have plagued the process. Here, we perform a qualitative assessment of the compliance of GRAS panels with basic elements of FDA’s guidance. We assessed 403 GRAS notices filed by FDA between 2015 and 2020 and identified whether a GRAS panel was convened and by whom, its members, affiliations, and relationships between panelists and panel conveners. Then, we compared FDA’s recommendations against the information included in the notices voluntarily submitted by manufacturers. We found no evidence that GRAS panels have adhered to FDA’s guidance. Panels are populated from a very small pool of professionals; we found that seven panel members alone occupied almost half of all available panel positions and that they often serve together. Against guidance recommendations, ad-hoc panels have been substituted by panels with recurring members in hired consulting firms’ payroll. The widespread persistence of conflicts of interest, appearance of conflict and bias in GRAS determinations continue to put the health of Americans at risk and undermine confidence in the safety of food ingredients in the US market. FDA should require notice for all GRAS determinations including how the financial conflicts of interest of those who make these determinations are minimized.

FEMA GRAS assessment of natural flavor complexes: Sage oil, Orris Root Extract and Tagetes Oil and related flavoring ingredients.

In recent years, the Expert Panel of the Flavor and Extract Manufacturers Association (FEMA) has conducted a program to re-evaluate the safety of natural flavor complexes (NFCs) used as flavor ingredients. This publication, twelfth in the series, details the re-evaluation of NFCs whose constituent profiles are characterized by alicyclic or linear ketones. In its re-evaluation, the Expert Panel applies a scientific constituent-based procedure for the safety evaluation of NFCs in commerce using a congeneric group approach. Estimated intakes of each congeneric group of the NFC are evaluated using the well-established and conservative Threshold of Toxicological Concern (TTC) approach. In addition, studies on the toxicity and genotoxicity of members of the congeneric groups and the NFCs under evaluation are reviewed. The scope of the safety evaluation of the NFCs contained herein does not include added use in dietary supplements or any products other than food. Thirteen (13) NFCs derived from the Boronia, Cinnamomum, Thuja, Ruta, Salvia, Tagetes, Hyssopus, Iris, Perilla and Artemisia genera are affirmed as generally recognized as safe (GRAS) under conditions of their intended use as flavor ingredients based on an evaluation of each NFC and the constituents and congeneric groups therein.